Recombinant PRC2 complex

Expressed In

Baculovirus

Protein Species

Human

Cat No : 31387,31887

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Validation Data Gallery

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  Recombinant PRC2 Complex gel. PRC2 Complex was run on a 10% SDS-PAGE gel and stained with Coomassie blue. MW as follows: EZH2: 87 kDa. SUZ12: 83 kDa. EED: 50.2 kDa. RbAp46: 47.8 kDa. RbAp48: 47.7 kDa. Purity: > 90%

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  HTRF assay for PRC2 Complex activity 1 uM H3 (16-37) peptide was incubated with different concentrations of PRC2 Complex in a 10 ul reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hours, then 10 ul of H3K27me1 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at RT. HTRF assay was used for detection.

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  HTRF assay for PRC2 Complex activity 1 uM H3 (16-37) peptide was incubated with different concentrations of PRC2 Complex in a 10 ul reaction system containing 50 mM Tris-HCl pH 8.6, 0.02% Triton X-100, 2 mM MgCl2, 1 mM TCEP and 50 uM SAM for 3 hours, then 10 ul of H3K27me1 antibody and SA-XL665 mixture (each 1:100 dilution in HTRF Detection Buffer) was added to each reaction system and incubated for 30 min. All the operations and reactions were performed at RT. HTRF assay was used for detection.

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Product Information

• Expressed In: Baculovirus
• Protein Species: Human

Contents

Recombinant PRC2 Complex is supplied in 25 mM HEPES-NaOH pH 7.5, 300 mM NaCl, 5% glycerol, 0.04% Triton X-100, and 0.5 mM TCEP.

Background

PRC2 (Polycomb Repressive Complex 2) is one of the two classes Polycomb-group, or PcG proteins (the other being PRC1) that are important epigenetic determinants of stem cell identity. They play an important role in long-term epigenetic silencing of genes during cell fate determination and differentiation. PRC2 functions as a repressor of chromatin. PRC2 is required to target recruitment to specific DNA sequences (termed Polycomb Response Elements or PREs) of genomic regions to be silenced. Once associated with chromatin, the PRC2 subunit EZH2 has histone methyltransferase activity that catalyzes the trimethylation of histone H3 at lysine 27. H3K27me3 is well established as a hallmark of regions of repressed chromatin. Trimethylation of lysine 27 leads to the recruitment of PRC1 through the binding of H3K27me3 by chromodomain-containing proteins in PRC1. PRC1 is responsible for long-term gene silencing after cellular differentiation.

Application Notes

This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product. All available data for a given product is shown on the lot-specific Technical Data Sheet.

Protein Details

Recombinant PRC2 Complex that includes full length EZH2, SUZ12, EED and RbAp46/48 (accession numbers NP_001190176.1, NP_056170, NP_003788.2, NP_002884.1, and NP_005601.1, respectively) was expressed in Sf9 and contains an N-terminal FLAG tag at the N-terminus of EZH2. The molecular weights of expressed EZH2, SUZ12, EED and RbAp46/48 are 87 kDa, 83 kDa, 50.2 kDa, 47.8 kDa and 47.7 kDa, respectively.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is guaranteed for 6 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.